GSK126对犬乳腺肿瘤细胞上皮间质转化的影响

doi:10.11843/j.issn.0366-6964.2023.04.035

摘要/Abstract

摘要： 探讨Zeste基因增强子同源物2（enhancer of Zeste homolog 2，EZH2）抑制剂GSK126对犬乳腺肿瘤细胞增殖、迁移、凋亡及上皮-间质转化（epithelial-mesenchymal transition，EMT）的影响。取对数生长期的犬乳腺肿瘤细胞（canine mammary tumor cells，CHMm cells），将不同浓度的EZH2抑制剂GSK126（0、10、20、40 μmol·L^-1）作用于体外培养CHMm细胞后，培养48 h，采用四甲基偶氮唑盐比色法（MTT）和细胞克隆形成试验检测细胞增殖能力，Transwell法检测细胞迁移能力变化，Annexin V-FITC/PI流式细胞仪检测细胞凋亡能力，qRT-PCR和Western blot方法检测CHMm细胞中凋亡相关因子Bcl-2和Bax、上皮间质标志物E-cadherin、N-cadherin、Vimentin、EZH2 mRNA和蛋白相对表达量。结果显示：与对照组相比，GSK126对CHMm细胞的增殖及迁移具有明显抑制作用，呈明显的剂量依赖性；与对照组相比，不同浓度GSK126处理组的凋亡率明显升高；GSK126处理后促凋亡因子Bax、上皮标志物E-cadherin mRNA和蛋白相对表达量均显著升高，抗凋亡因子Bcl-2、间质标志物N-cadherin、Vimentin和EZH2 mRNA和蛋白相对表达量均明显降低。EZH2抑制剂GSK126可通过抑制犬乳腺肿瘤细胞的增殖、迁移、EMT和诱导细胞凋亡，从而抑制犬乳腺肿瘤细胞发展进程，为犬乳腺肿瘤治疗提供理论基础。

关键词: GSK126, 犬乳腺肿瘤, 细胞增殖, 凋亡, 上皮-间质转化

Abstract: This study was conducted to investigate the effect of enhancer of Zeste homolog 2 (EZH2) inhibitor GSK126 on the proliferation, migration, apoptosis and epithelial-mesenchymal transition (EMT) of canine mammary tumor cells. Canine mammary tumor cells (CHMm cells) in logarithmic growth phase were collected, and different concentrations of GSK126 (0, 10, 20, 40 μmol·L^-1) were used to culture CHMm cells. After 48 h, tetramethylazolium salt colorimetric method (MTT) and cell clone formation assay were detected for the proliferation ability of cells. Transwell assay was performed to evaluate the migration ability. The cell apoptosis was detected by Annexin V-FITC/PI flow cytometry, and the relative expression of apoptosis-related factors Bcl-2 and Bax, epithelial-mesenchymal markers E-cadherin, N-cadherin, Vimentin, EZH2 were determined by qRT-PCR and Western blot method. After treated with GSK126 for 48 h, the proliferation and migration of CHMm cells were inhibited in a concentration-dependent manner. The apoptosis rate of the GSK126 treatment group was significantly increased. The relative expression of pro-apoptotic factor Bax, epithelial marker E-cadherin mRNA and protein compared with control group was significantly increased as the concentration of GSK126 treatment group, in contrast, anti-apoptotic factor Bcl-2, mesenchymal marker N-cadherin, Vimentin and EZH2 mRNA and protein was significantly reduced. EZH2 inhibitor GSK126 may inhibit the proliferation, migration, EMT and induce apoptosis of canine mammary tumor cells, which provides a theoretical basis for the treatment of canine mammary tumor cells.

Key words: GSK126, canine mammary tumor, cell proliferation, apoptosis, epithelial-mesenchymal transition

中图分类号:

任晓丽, 范玉营, 皇甫和平, 刘云, 石冬梅. GSK126对犬乳腺肿瘤细胞上皮间质转化的影响[J]. 畜牧兽医学报, 2023, 54(4): 1721-1729.

REN Xiaoli, FAN Yuying, HUANGFU Heping, LIU Yun, SHI Dongmei. Effect of GSK126 on Epithelial-mesenchymal Transition of Canine Mammary Tumor Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1721-1729.

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